Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing 400014, PR China; Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing 400014, PR China; Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing 400014, PR China; National Clinical Research Center for Child Health and Disorders, Chongqing 400014, PR China; China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing 400014, PR China; Chongqing Key Laboratory of Pediatrics, Chongqing 400014, PR China.
Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing 400014, PR China; Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing 400014, PR China; National Clinical Research Center for Child Health and Disorders, Chongqing 400014, PR China; China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing 400014, PR China; Chongqing Key Laboratory of Pediatrics, Chongqing 400014, PR China.
J Hazard Mater. 2021 Mar 15;406:124316. doi: 10.1016/j.jhazmat.2020.124316. Epub 2020 Oct 19.
Di(2-ethylhexyl) phthalate (DEHP) is a widely-used plasticizer and has long been recognized as an endocrine-disrupting chemical with male reproductive toxicities. DEHP exposure at the prepubertal stage may lead to extensive testicular injury. However, the underlying mechanisms remain to be elucidated. In the present study, we gavaged male C57BL/6 mice with different concentrations of DEHP (0, 250, and 500 mg/kg-bw·d) from postnatal day 22-35, and exposed TM3 Leydig cells with 0, 100, 200, 300, and 400 μM of MEHP (bioactive metabolite of DEHP) for 12-48 h. RNA sequencing was performed both in testicular tissue and TM3 cells. The results showed that DEHP disrupts testicular development and reduces serum testosterone levels in male prepubertal mice. Bioinformatic analysis and experimental verification have revealed that DEHP/MEHP induces cell cycle arrest in TM3 cells and increases apoptosis both in vivo and in vitro. Furthermore, the p53 signaling pathway was found to be activated upon DEHP/MEHP treatment. The inhibition of p53 by pifithrin-α significantly reduced MEHP-induced injuries in TM3 cells. Cumulatively, these findings revealed the involvement of the p53 signaling pathway in DEHP-induced prepubertal testicular injury by promoting cell apoptosis and inhibiting cell proliferation of Leydig cells.
邻苯二甲酸二（2-乙基己基）酯（DEHP）是一种广泛使用的增塑剂，长期以来一直被认为是一种具有男性生殖毒性的内分泌干扰化学物质。青春期前阶段的 DEHP 暴露可能导致广泛的睾丸损伤。然而，其潜在机制仍有待阐明。在本研究中，我们从出生后第 22-35 天开始给雄性 C57BL/6 小鼠灌胃不同浓度的 DEHP（0、250 和 500mg/kg-bw·d），并用 0、100、200、300 和 400μM 的 MEHP（DEHP 的生物活性代谢物）处理 TM3 间质细胞 12-48h。在睾丸组织和 TM3 细胞中均进行了 RNA 测序。结果表明，DEHP 破坏青春期前雄性小鼠的睾丸发育并降低血清睾酮水平。生物信息学分析和实验验证表明，DEHP/MEHP 诱导 TM3 细胞细胞周期停滞，并在体内和体外增加细胞凋亡。此外，发现 DEHP/MEHP 处理后激活了 p53 信号通路。pifithrin-α 抑制 p53 显著减少了 MEHP 诱导的 TM3 细胞损伤。综上所述，这些发现揭示了 p53 信号通路通过促进间质细胞凋亡和抑制细胞增殖参与 DEHP 诱导的青春期前睾丸损伤。
